Method for treating cytokine mediated hepatic injury

ABSTRACT

A method of modulating cytokine mediated hepatic injury by administering compound-D SEQ ID NO: 1 to a mammal. A concentration of the compound in the range of about 0.5 mg/kg to about 20 mg/kg in a physiologically acceptable formulation blocks a cytokine cascade. A therapeutic method of modulating cytokine mediated acute inflammatory, trauma induced and toxin induced hepatic injury, particularly via tumor necrosis factor modulation, is thus disclosed.

[0001] This application claims the benefit of U.S. application Ser. No.60/238,991, filed Oct. 10, 2000.

FIELD OF THE INVENTION

[0002] The invention relates to the use of compounds to attenuate orprevent cytokine mediated hepatic injury.

BACKGROUND

[0003] Hepatic injury can be caused by a number of different agentsincluding viruses such as Hepatitis A, B, C, D and E, both gram positiveand gram negative bacteria, chemical agents such as ethanol, carbontetrachloride and lead, and by physical trauma resulting in ischemia(ischemic hepatitis) injuries as can occur in right-sided congestiveheart failure. It is now believed that all of these types of hepaticinjury are caused at least in part by the liver's inflammatory orcytokine response to these agents. The inflammatory response of theliver results in the overexpression of a cascade of inflammatory/acutephase cytokines, such as interleukin-1 (IL-1), tumor necrosis factor(TNF), IL-6, IL-8 and transforming growth factor beta (TGFβ). It is nowbelieved that it is the cascade of these cytokines which is the ultimatecause of much of the hepatic injury resulting from these agents. Thus,there is a need for a therapeutic agent which can be useful inalleviating or modulating the inflammatory response associated withliver disease or injury.

SUMMARY OF THE INVENTION

[0004] The present invention fills this need by providing a method oftreating or preventing a cytokine mediated hepatic injury in a mammalcomprised of administering a pharmaceutically effective amount of apeptide having the sequenceTyr-D-Leu-Phe-Ala-Asp-Val-Ala-Ser-Thr-Ile-Gly-Asp-Phe-Phe-His-Ser-Ile-NH₂SEQ ID NO: 1, hereinafter referred to as compound D, to said mammal. Thehepatic injury can be an acute inflammatory reaction, as a result of aviral or bacterial infection or a chemical agent such as ethanol, lead,carbon tetrachloride or acetaminophen, or from trauma resulting inischemia or reperfusion injury in the liver.

[0005] The present invention is also directed to a method of treating aviral or bacterial infection-related hepatic damage in a mammalcomprised of administering a pharmaceutically effective amount ofcompound D SEQ ID NO: 1 to said mammal.

[0006] The present invention is also directed to a method of treatingalcohol induced liver injury in a mammal comprised of administering apharmaceutically effective amount of compound D SEQ ID NO: 1 to saidmammal.

[0007] Preferably, compound D SEQ ID NO: 1 is administered in apharmaceutical composition at a dosage of from about 0.5 mg/kg to about20 mg/kg per body weight of the mammal.

[0008] Preferably, the mammal is a human.

DETAILED DESCRIPTION

[0009] A compound used to treat cytokine-mediated hepatic injury is apeptide having the sequenceTyr-D-Leu-Phe-Ala-Asp-Val-Ala-Ser-Thr-Ile-Gly-Asp-Phe-Phe-His-Ser-Ile-NH₂SEQ ID NO: 1, hereinafter referred to compound-D. The peptide may beproduced by a number of methods, such as using an automated peptidesynthesizer, through recombinant molecular techniques, or isolated froma naturally occurring source, as is known to one skilled in the art.Compound-D SEQ ID NO: 1 has a molecular weight of 1,902 daltons.Compound-D SEQ ID NO: 1 is insoluble in water or saline, but may besolubilized by adding 100 μM of a solution comprised of ethanol,propylene glycol, and 1 N NaOH in a 1:1:1 ratio, with sterilephysiological saline then used to obtain the appropriate concentration.The initial alkaline pH is adjusted to 7.4 with 1 N HCl.

[0010] Compound-D SEQ ID NO: 1 that has been solubilized may beadministered by parenteral means, for example, by intravenous injection.For administration into a mammal, a dose of about 1-20 milligrams perkilogram (mg/kg) is useful. For administration into a tissue or organpreservation solution, a concentration of about 100 μM is useful.

[0011] Compound-D SEQ ID NO: 1 may be administered directly into amammal, either alone or in combination with other substances.

[0012] The above agent is administered to a mammal to modulate cytokineactivation by blocking one or more steps in the cytokine cascade. Theagent may be formulated for administration in an aqueous based liquidsuch as phosphate buffered saline to form an emulsion, or may beformulated in an organic liquid such as dimethylsulfoxide to form asolution. The solution or emulsion may be administered by any route, butit is preferably administered parenterally such as by intravenous,intramuscular, intradermal or intraperitoneal injections. A preferreddose is in the range of about 0.5-20 mg of compound-D SEQ ID NO: 1 perkg of body weight of the mammal. The time of administration of the agentis preferably prior to initiation of cytokine activation. However, theagent may be administered concurrently with another agent that inducescytokine activation or even subsequent to an agent that induces cytokineactivation and still produce a protective effect.

[0013] Administration of compound-D SEQ ID NO: 1 should be continued ona daily basis until hepatic function returns to normal and is maintainedat normal levels, preferably for at least one to two days. Hepaticinjury can be determined by elevated levels of hepatic enzymes, as wellas by depressed albumin levels (less than about 35 g/liter). Hepaticfunction is routinely monitored by quantitating serum levels of hepaticenzymes such as alanine aminotransferase (ALT) (normal <35 U/L),aspartate aminotransferase (AST) (normal <30 U/L), alkaline phosphatase(ALP) (normal ≦100 U/L) and gamma glutamyltransferase (GGT) (normal ≦45U/L for males, ≦30 U/L for females), as well as bilirubin, bothconjugated (normal ≦0.2 mg/deciliter) and total (normal ≦1.0mg/deciliter) bilirubin. Compound-D SEQ ID NO: 1 modulation ofhepatocyte cytokine activation may be used therapeutically in a varietyof hepatic injury processes. As used herein, the term hepatic injurybroadly encompasses all types of injury such as hepatic trauma, physicaland/or chemical insult, stress, inflammation, toxicity, disease and soon. For example, the inventive agents can be used in treating hepaticinjury due to alcoholic liver disease, acetaminophen toxicity, cadmiumtoxicity, lead poisoning, bacteremia due to, for example, Staphylococcusspecies, Streptococcus species, Neisseria species, Salmonella species,Shigella species, Escherichia coli, Clostridium perfringens, Klebsiellaspecies, Proteus species, Enterobacter species, Bacteroides species,Brucella species, Francisella tularensis, Listeria monocytogenes,Acinetobacter species, Streptobacillus moniliformis, Vibrio species,Helicobacterpylori, Pseudomonas species, Haemophilus species, Bordetellapertussis, viral infections due to, for example, influenza viruses,adenoviruses, paramyxoviruses, rubella viruses, polioviruses, hepatitisviruses, herpesviruses, rabies viruses, human immunodeficiency virusesand papilloma viruses, as well as trauma, ischemia reperfusion injuryand metabolic liver disease.

[0014] While the specific mechanism of action of compound-D SEQ ID NO: 1on the modulation of cytokine mediated hepatic injury such as acuteinflammatory reactions, trauma and toxin induced biological responses isunknown, these agents exhibit a specific and reproducible effect ondecreasing hepatotoxicity.

[0015] A treatment for attenuating and/or preventing cytokine mediatedacute inflammatory, trauma induced and toxin induced hepatic injury isthus disclosed. Compound-D SEQ ID NO: 1, administered at a concentrationof about 0.5 mg/kg to about 20 mg/kg, inhibits hepatic injury and resultin decreased lethality of an injured animal.

[0016] It should be understood that the embodiments of the presentinvention shown and described in the specification are only preferredembodiments of the inventors who are skilled in the art and thus are notlimiting in any way. Therefore various changes, modifications oralterations to these embodiments may be made or resorted to withoutdeparting from the spirit of the invention and the scope of thefollowing claims.

1 1 1 17 PRT Artificial Sequence MOD_RES (1)...(17) Xaa = D-Leu;artificial sequence is completely synthesized 1 Tyr Xaa Phe Ala Asp ValAla Ser Thr Ile Gly Asp Phe Phe His Ser Ile 1 5 10 15

What is claimed is:
 1. A method of modulating a cytokine mediatedhepatic injury response in a mammal comprising administering compound-DSEQ ID NO: 1 to the mammal in a pharmaceutically acceptable formulation.2. The method of claim 1 wherein said compound is administered prior tosaid response.
 3. The method of claim 1 wherein said compound isadministered subsequent to said response.
 4. The method of claim 1wherein said compound is administered substantially concurrently withsaid response.
 5. The method of claim 1 wherein said compound isadministered in the formulation selected from the group consisting of asolution, an emulsion and a suspension.
 6. The method of claim 1 whereinsaid compound is administered parenterally.
 7. The method of claim 1wherein said compound is administered at a concentration in the range ofabout 0.5 mg/kg to about 20 mg/kg.
 8. A method for treating hepaticinjury in a mammal by a chemical toxin comprising administering apharmaceutically effective concentration of compound-D SEQ ID NO:
 1. 9.The method of claim 8 wherein the chemical toxin is selected from thegroup consisting of ethanol, lead, cadmium, carbon tetrachloride, andacetaminophen.
 10. A method for treating a bacterial or viral infectionrelated hepatic injury in a mammal comprising administering apharmaceutically effective concentration of compound-D SEQ ID NO:
 1. 11.The method of claim 10 wherein the bacterial or viral infection iscaused by an organism selected from the group consisting ofStaphylococcus species, Streptococcus species, Neisseria species,Salmonella species, Shigella species, Escherichia coli, Clostridiumperfringens, Klebsiella species, Proteus species, Enterobacter species,Bacteroides species, Brucella species, Francisella tularensis, Listeriamonocytogenes, Acinetobacter species, Streptobacillus moniliformis,Vibrio species, Helicobacter pylori, Pseudomonas species, Haemophilusspecies, Bordetella pertussis, influenza viruses, adenoviruses,paramyxoviruses, rubella viruses, polioviruses, hepatitis viruses,herpesviruses, rabies viruses, human immunodeficiency viruses andpapilloma viruses.